Brazilian Journal of Pharmaceutical Sciences Article http://dx.doi.org/10.1590/s2175-97902017000300260 Chemical compositions and antioxidant activity of Heracleum persicum essential oil Maryam Gharachorloo1*, Masoud Honarvar1, Shima Mardani2 1Department of Food Science and Technology, Science and Research Branch, Islamic Azad University, Tehran, Iran, 2Department of Chemical Engineering, Science and Research Branch, Islamic Azad University, Tehran, Iran In this study essential oil of the aerial parts of Heracleum persicum a spice widely used in Iran was isolated by conventional hydrodistillation (HD) and microwave-assisted hydrodistillation (MAHD) techniques. The extraction yield was determined and the chemical compositions of essential oils were identified by the application of gas chromatography/mass spectrometry (GC/MS). The antioxidant activity was determined by two different methods: 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging and oven test methods. Although the main compounds of essential oils by the both extraction methods were similar, the essential oil extracted by HD with lower extraction efficiency showed more diverse compounds. The evaluation of antioxidant activity of the essential oil measured by delay in sunflower oil oxidation indicated that the antioxidant activity was dependent on the concentration which increased when higher concentrations of the essential oils were applied. The results of DPPH radical assay also indicated that the percentage of inhibition increased with increasing of essential oil concentration and IC50 value for essential oil extracted by MAHD method was obtained 1.25 mg/mL. Therefore the Heracleum persicum essential oil might be recommended for use as a flavoring agent and a source of natural antioxidants in functional foods, formulation of the supplements and in medicinal due to numerous pharmacological activities. Keywords: Heracleum persicum/essential oil/chemical composition. Heracleum persicum/functional properties. Heracleum persicum/antioxidant activity. INTRODUCTION Among different methods for extraction of essential oils from spices, hydrodistillation is the common and most Heracleum persicum is a flowering plant and frequently used method (Morsy, 2015). However, in order native to Iran that commonly known as “Golpar”. It to reduce the extraction time, the operation costs, possibly belongs to Apiaceae family in the order of Apiales that improve the yield and the quality of the essential oils, contains about 300 genera and more than 3000 species. new approaches such as microwave-assisted extraction, Heracleum species probably originates from the Middle supercritical fluid extraction and ultrasound-assisted East, somewhere south of Caucasian, but has spread as an extraction have also been in consideration (Memarzadeh, ornamental plant up to Northern Europe. Heracleum genus Ghasemi Pirbalouti, Adibnejad, 2015). has 10 species in Iran (Asgarpanah et al., 2012). In this regards some advances in using microwave The ripe fruits, leaves and young stems of Heracleum have resulted in a number of techniques such as microwave persicum are used as antiseptic, carminative, digestive and steam diffusion (Farhat et al., 2011), microwave analgesia as well as flavoring in the Iranian folk medicine hydrodiffusion and gravity (Vianet al., 2008), microwave (Torbati et al., 2014). Also it has recently been shown to dry distillation (Tigrine Korjani, Meklati, Chemat, 2006) have antioxidant, anticonvulsant, anti-inflammatory and and solvent free microwave extraction (Lucchesi et immunemodulatory activities (Asgarpanah et al., 2012). al., 2007). Some recently studies (Golmakani, Rezaei, 2008; Perino Issartier et al., 2013; Stashenko, Jaramillo, *Correspondence: M. Gharachorloo. Faculty of Food Science and Tech- Martinez, 2004; Tigrine Kordjani, Meklati, Chemat, nology, Science and Reserch Branch, Islamic Azad University, Hesarak, Ashrafi Esfahani, Postal code: 1477893855 - Poonak, Tehran, Iran. E-mail: 2006) have successfully utilized a microwave oven for the [email protected] extraction of active components from medicinal plants. Braz. J. Pharm. Sci. 2017;53(3):e00260 Page 1 / 6 M. Gharachorloo, M. Honarvar, S. Mardani Therefore the aim of this research was the Chemical composition of essential oils comparison of the chemical compositions and antioxidant activity of Heracleum persicum essential oil extracted The chemical compositions of essential oils by conventional hydrodistillation (HD) and microwave- obtained from aerial parts of Heracleum persicum assisted hydrodistillation (MAHD) techniques. by using hydrodistillation and microwave-assisted hydrodistillation methods were identified by the MATERIALS AND METHODS application of gas chromatography/mass spectrometry (GC/MS). Samples were analyzed by using an Agilent Material Hp-6890 gas chromatograph (Agilent Technologies, USA) with a Hp-1 Methyl silicone capillary column (60 Dried aerial parts of Heracleum persicum a spice m×0.32 mm×0.25 µm film thickness) linked to Hp-5973 widely used in Iran were supplied from Tehran market. mass spectrometry detector. Oven temperature was set as The identity of the Heracleum was certificated by top 60 °C for 3 min initially and increased to 230 °C, at a rate experts from Herbarium of the Islamic Azad University, of 7 °C/min and subsequently held isothermal for 3min. Science and Research Branch. Aerial parts of Heracleum Injector temperature was set as 250 °C. Helium was used persicum were ground in a mill (Triplex, France), passed as carrier gas at a flow rate of 1 mL/min and 1 µL sample through a sieve of 0.5 mm mesh and the powder obtained was injected manually in the split mode. Ionization of was stored in plastic bags, in darkness at 4 °C. Refined sample compounds was performed in the ET mode (70 ev). sunflower seed oil without added synthetic antioxidant was The identification of compounds was performed by purchased from Behshahr Oil Company of Tehran. All the comparison of their mass spectral pattern and their linear chemicals used were of analytical grade, purchased from retention indices based on a homologous series of normal Merck Chemical Company of Germany. alkanes (C8–C20) with those of authentic references and the Wiley 257 mass spectra database. Isolation of essential oil Antioxidant activity Hydrodistillation Powder of aerial parts of Heracleum persicum were DPPH free radical scavenging assay hydrodistillated for 3 h using a Clevenger-type apparatus The free radical scavenging activities of essential according to the standard procedure. The essential oil oils were measured using 1,1-diphenyl-2-picrylhydrazyl volume was measured directly in the extraction burette. (DPPH) as described by El Ouariachi et al. (2014). The obtained essential oils were dried with anhydrous Antioxidants react with the stable free radical DPPH sodium sulphate and then stored in a sealed dark vials at (deep violet color) and convert it to 1,1-diphenyl- 4°C until further analysis. Yield percentage was calculated 2-picrylhydrazine with discoloration. Various as volume (mL) of essential oil per 100 g of plant dry concentrations of essential oils were added to DPPH matter (Jordán et al., 2009). radical solution in ethanol. The mixture was strongly shaken and left to stand at room temperature for 30 min Microwave-assisted hydrodistillation (MAHD) in the dark. The absorbance was measured at 517 nm by A domestic microwave oven (micro SYNTH, 2450 using a spectrophotometer (Cary 50 Scan UV-Visible MHz) was modified for MAHD operation. The interior Spectrophotometer). The radical scavenging activity was cavity of the microwave oven was 29 × 37 × 40 cm. Flat expressed as percentage of inhibition (I%) according to bottom flask having a capacity of 500 mL was placed the following equation: inside the cavity for the MAHD experiments. 20gr of powdered samples soaked in 100 mL of distilled water for I(%)=[1-(Asample / A control)]× 100 30 minutes were placed in a 500 mL flask. The flask was setup within the microwave oven cavity and a condenser where Acontrol is the absorbance of the control reaction and was used on the top, outside the oven. The microwave oven Asample is the absorbance of the test compound. was operated at 600 power level for a period of 30 min The sample concentration providing 50% inhibition which no more essential oil was obtained. The extracted (IC50) was calculated from the graph of inhibition essential oils were collected, dehydrated with anhydrous percentage against sample concentration. Tests were sodium sulfate and kept in a sealed dark vials at 4 °C carried out in triplicate. Ascorbic acid was used as a (Karakaya et al., 2014). positive control. Page 2 / 6 Braz. J. Pharm. Sci. 2017;53(3):e00260 Chemical compositions and antioxidant activity of Heracleum persicum essential oil Oven test method started at much earlier time than HD method (5min and The essential oil extracted by MAHD method was 35 min respectively). The process continued until no more added to 100 g sunflower seed oil at the concentrations of essential oil was obtained. Full extraction of essential oils 0.02, 0.04, 0.06, 0.08 and 0.1% (w/w) to examine their was achieved within the first 30 min of operation with antioxidant activity. The antioxidant activity of extracts MAHD method. In the case of HD method, a time period was compared with tertiary butylhydroquinone (TBHQ) of at least 3 h was necessary for such purpose. at the concentration of 0.01% (w/w). All the samples It seems that it is due to the more efficient heat